SBIR-STTR Award

Improved supporting technologies for imaging of molecular and supramolecular structures within cells are needed to facilitate cell biology research, and are sought by the National Institute of General Medical Sciences (specifically, its Division of Cell Biology and Biophysics). Multiscale imaging, using cryo-electron tomography (cryo-ET) on supramolecular structures and single molecules, has proven in recent years to be a unique and invaluable method for high-throughput characterization of the dynamic 3D architecture of cells. Electron microscopy (EM) grids, used as substrates for supporting the biological and biomolecular specimens being imaged, are a critical component associated with this imaging method. New EM grid technology that decreases sample preparation cost and time, improves sample generation from culturing to freezing for cryo-ET, and increases imaging quality will allow researchers to more efficiently explore cellular architecture, at higher throughput. Synkera proposes a novel ceramic EM grid that features an integrated thin support film that is highly compatible with cell culturing, light microscopy and cryo-ET. The grids will facilitate high-throughput, multiscale imaging of sub-cellular architecture and offer key advantages over state-of-the art products. The grids are also expected to be a competitive alternative in many other EM and culturing applications. Phase I work will demonstrate feasibility of the proposed fabrication method, based on micromachined nanoporous ceramic. Phase I will also demonstrate compatibility of the proposed EM grids with cell culturing, light microscopy and cryo-EM. EM grids will be fabricated with two different integrated support film options. These grids will be compared to traditional EM grid products (gold grids with holey carbon thin film supports) in order to demonstrate greater performance for multiscale cellular and molecular imaging. At least three academic partners will aide in demonstrating these capabilities.

Public Health Relevance:
The project addresses imaging of molecules and cells via cryo-electron tomography (cryo-ET). Specifically, the target application is multiscale imaging via optical microscopy and cryo-ET of cellular, supramolecular and single-molecule structures, for generating 3D models of sub-cellular architecture. The development of a novel class of ceramic-based electron microscopy grids that facilitate this multiscale imaging is proposed. The proposed technology will offer greater capability over state-of-the-art products and help further streamline multiscale cellular imaging by simplifying the specimen preparation process and yielding superior imaging performance.

Thesaurus Terms:
3d Modeling; Address; Adhesions; Architecture; Arts; Au Element; Biological; Biophysics; C Element; Carbon; Cell Culture Techniques; Cells; Cellular Biology; Ceramic; Ceramics; Colorado; Cryo-Electron Microscopy; Cryoelectron Microscopy; Cultured Cells; Data; Data Quality; Development; Dimensions; Electron Cryomicroscopy; Electron Microscopy; Engineering / Architecture; Evaluation; Film; Foundations; Freezing; Generations; Goals; Gold; Government; Image; Imaging Technology; Investigators; Laboratories; Maps; Marketing; Methods; Microscopy; Molecular; Nigms; National Institute Of General Medical Sciences; Optics; Performance; Persons; Phase; Preparation; Process; Relative; Relative (Related Person); Research; Research Personnel; Research Specimen; Researchers; Sampling; Science; Solid; Specimen; Staging; Structure; Technology; Time; Universities; Work; Base; Bioimaging; Bioimaging/Biomedical Imaging; Biomedical Imaging; Cell Biology; Cell Imaging; Cellular Imaging; Cost; Cryoem; Electron Tomography; Imaging; Imaging Modality; Improved; Light Microscopy; Meetings; Molecular Imaging; Molecular/Cellular Imaging; Novel; Professor; Prototype; Public Health Relevance; Single Molecule; Three-Dimensional Modeling; Tomography

Improved supporting technologies for imaging of molecular and supramolecular structures within cells are needed to facilitate cell biology research, and are sought by the National Institute of General Medical Sciences (specifically, its Division of Cell Biology and Biophysics). Multiscale imaging, using cryo-electron tomography (cryo-ET) on supramolecular structures and single molecules, has proven in recent years to be a unique and invaluable method for highthroughput characterization of the dynamic 3D architecture of cells. Electron microscopy (EM) grids, used as substrates for supporting the biological and biomolecular specimens being imaged, are a critical component associated with this imaging method. New EM grid technology that decreases sample preparation cost and time, improves sample generation from culturing to freezing for cryo-ET, and increases imaging quality will allow researchers to more efficiently explore cellular architecture, at higher throughput. Synkera proposes a novel class of ceramic EM grids that feature an integrated thin support film that is highly compatible with cell culturing, light microscopy and cryo-ET. The grids will facilitate high-throughput, multiscale imaging of sub-cellular architecture and offer key advantages over state-of-the art products. The grids are also expected to be a competitive alternative in many other EM and culturing applications a 6-month Phase I project successfully demonstrated feasibility of the proposed grids architecture, as well as their potential in culturing, cryo-ET and multiscale imaging. Phase II aims to build on this success by further developing fabrication processes and ceramic EM grids designs, to fully realize the potential for multiscale imaging in functional prototypes. At least four academic partners will aide in this development process. The ultimate goal of the proposed project is a complete line of ceramic EM grids for a broad range of EM applications, from bioimaging to materials characterization.

Public Health Relevance:
The project addresses imaging of molecules and cells via cryo-electron tomography (cryo-ET). Specifically, the target application is multiscale imaging via optical microscopy and cryo-ET of cellular, supramolecular and single-molecule structures, for generating 3D models of sub-cellular architecture. The development of a novel class of ceramic-based electron microscopy grids that facilitate this multiscale imaging is proposed. The proposed technology will offer greater capability over state-of-the- art products and help further streamline multiscale cellular imaging by simplifying the specimen preparation process and yielding superior imaging performance.

Thesaurus Terms:
3d Modeling;Address;Architecture;Articulation;Automation;Biologic Sciences;Biological;Biological Sciences;Biophysics;Case Study;Cell Components;Cell Culture Techniques;Cell Structure;Cells;Cellular Structures;Cellular Biology;Ceramics;Colorado;Communities;Cryo-Electron Microscopy;Cryoelectron Microscopy;Development;Educational Mainstreaming;Electron Cryomicroscopy;Electron Microscopy;Engineering / Architecture;Evaluation;Film;Freezing;Generations;Generic Drugs;Goals;Image;Imaging Technology;Investigators;Joints;Laboratories;Life Sciences;Mainstreaming;Mainstreaming (Education);Methods;Microscopy;Nigms;National Institute Of General Medical Sciences;Nonproprietary Drugs;Optics;Performance;Phase;Preparation;Process;Protocol;Protocols Documentation;Publications;Research;Research Institute;Research Personnel;Research Specimen;Researchers;Sampling;Scientific Publication;Specimen;Structure;Techniques;Technology;Time;Universities;Achievement Mainstream Education;Base;Bioimaging;Biomedical Imaging;Case Report;Cell Biology;Cell Culture;Cell Imaging;Cellular Imaging;Comparative;Cost;Cryoem;Design;Designing;Developmental;Electron Tomography;Generic;Imaging;Imaging Method;Imaging Modality;Improved;Light Microscopy;Meetings;Molecular Imaging;Molecular Scale;Novel;Optical;Prototype;Scale Up;Single Molecule;Success;Three-Dimensional Modeling